title: Adenoviral-mediated gene transfer into bone marrow: an effective surgical technique in rat.
abstract: BACKGROUND
  The role of transforming growth factor-beta 1 (TGF-β₁) in the onset of bone marrow fibrosis has been confirmed in some animal models. To further understand the genetic expression of some myeloproliferative disorders affecting marrow stem cells, however, it is necessary to develop a specific and reliable procedure to deliver modified adenoviral vectors into the bone marrow cavity. The aim of this paper is to report a surgical technique designed to deliver an adenoviral vector-mediated gene expressing TGF-β₁ into the bone marrow of rat femurs.
  
  
  METHODS
  Forty-two Sprague-Dawley rats were used in the study. Rat femurs were exposed and the compact and trabecular bones at the proximal head removed. An intrabone marrow injection of a mutated TGF-β₁ adenoviral vector, a null adenoviral vector, or PBS was delivered into the bone. Three groups were accounted (n = 14 per group): fibrogenic and positive and negative controls. The quality of the surgical entrance was assessed by means of computerized tomography and histological changes were assessed by histochemistry. The concentration of TGF-β₁ in the bone marrow was determined by ELISA.
  
  
  RESULTS
  The surgical technique was conducted under ideal timing (approx. 10 min) and no surgical or postsurgical complications were observed. Computerized tomography revealed no changes in the bone tissue and a clean entrance was delimited through the bone to the bone marrow. HE and Masson's trichrome staining indicated highly fibrotic areas in the profibrotic group and bone marrow lavage reported a significantly higher concentration of TGF-β₁ (p < 0.05) in that same group.
  
  
  CONCLUSIONS
  The present study confirmed that the proposed surgical technique is an effective method to deliver adenoviral vectors into the femoral bone marrow to investigate the physiopathology of bone marrow fibrosis in rats.
authors.affiliation: Department of Pathology and Microbiology, Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, P.E.I., Canada. jrodriguez@upei.ca
authors.forename: J C
authors.initials: JC
authors.lastname: Rodriguez-Lecompte
authors.forename: G A
authors.initials: GA
authors.lastname: Romero-Perez
authors.forename: G
authors.initials: G
authors.lastname: Ramirez-Yañez
authors.forename: K
authors.initials: K
authors.lastname: Ask
authors.forename: J
authors.initials: J
authors.lastname: Gauldie
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001853:Bone Marrow / Q000000981:diagnostic imaging / Q000378:metabolism* / Q000601:surgery*; D005269:Femur / Q000000981:diagnostic imaging / Q000378:metabolism / Q000601:surgery; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D055728:Primary Myelofibrosis / Q000235:genetics / Q000601:surgery / Q000628:therapy; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014057:Tomography, X-Ray Computed; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D053773:Transforming Growth Factor beta1